Ferula species have occupied great value in traditional medicine in various countries. The aim of the present paper was to assess for the first time to the phytochemical profile of Ferula tunetana Pomel ex Batt. (F. tunetana) seed and leaf extracts using HPLC-MS/MS, as well as to evaluate their antioxidant and anti-α-amylase activities. Therefore, total phenolic and flavonoid amounts of the studied extracts were determined and 38 compounds were detected and quantified (phenolic acids, flavonoids, non-phenolic acid and stilbenes). In addition of their antioxidant potential against 1,1-diphenyl-2-picrylhydrazyl (DPPH) and Ferric Reducing Antioxidant Power (FRAP) assays, in vitro and in silico studies were adopted to evaluate α-amylase inhibitory properties of F. tunetana extracts. Ethyl acetate (EtOAc) seed extract showed the highest total phenolic content, antioxidant potential (IC50 = 19.00 ± 0.37 μg/mL) in terms of free radical scavenging, also for α-amylase inhibition activity with IC50 values of 17.39 ± 0.92 μg/mL. The molecular docking analysis and the drug-likeness prediction of the major selected compounds explained the significant anti-α-amylase activity of the studied extracts. The obtained analysis and results suggest that F. tunetana extracts could be promising candidates to develop a new generation of efficient and especially safe antidiabetic treatments. Display omitted •38 phytochemicals were quantified in Ferula tunetana extracts by HPLC-MS/MS method.•Total phenolic and flavonoid contents in seed and leaf extracts were estimated.•The studied extracts exhibited good anti-α-amylase and antioxidant activities.•The molecular docking of main components validated the strong antidiabetic effects.•The drug-likeness proprieties of the major compounds were predicted.